Naphthalenetetrayltetrakis(sulfonylimino)-aryl multicarboxylic acids and salts thereof

ABSTRACT

Naphthalenetetrayltetrakis(sulfonylimino)-aryl multicarboxylic acids and salts thereof useful as complement inhibitors.

BACKGROUND OF THE INVENTION

The present invention resides in the concept of certainnaphthalenetetrayltetrakis(sulfonylimino)-aryl multicarboxylic acids andsalts thereof as novel compounds and their use as inhibitors of thecomplement system of warm-blooded animals.

The term "complement" refers to a complex group of proteins in bodyfluids that, working together with antibodies or other factors, play animportant role as mediators of immune, allergic, immunochemical and/orimmunopathological reactions. The reactions in which complementparticipates takes place in blood serum or in other body fluids, andhence are considered to be humoral reactions.

With regard to human blood, there are at present more than 11 proteinsin the complement system. These complement proteins are designated bythe letter C and by number: C1, C2, C3 and so on up to C9. Thecomplement protein C1 is actually an assembly of subunits designatedC1q, C1r and C1s. The numbers assigned to the complement proteinsreflect the sequence in which they become active, with the exception ofcomplement protein C4, which reacts after C1 and before C2. Thenumerical assignments for the proteins in the complement system weremade before the reaction sequence was fully understood. A more detaileddiscussion of the complement system and its role in the body processescan be found in, for example, Bull. World Health Org., 39, 935-938(1968); Ann. Rev. Medicine, 19, 1-24 (1968); The John Hopkins Med. J.,128, 57-74 (1971); Harvey Lectures, 66, 75-104 (1972); The New EnglandJournal of Medicine, 287, 452-454; 489-495; 545-549; 592-596; 642-646(1972); Scientific American, 229, (No. 5), 54-66 (1973); FederationProceedings, 32, 134-137 (1973); Medical World News, Oct. 11, 1974, pp.53-66; J. Allergy Clin. Immunol., 53, 298-302 (1974); Cold Spring HarborConf. Cell Proliferation 2/Proteases Biol. Control/229-241 (1975); Ann.Review of Biochemistry, 44, 697 (1975); Complement in Clinical Medicine,Disease-a-Month, (1975); Complement, Scope, December 1975; Annals ofInternal Medicine, 84, 580-593 (1976); "Complement: Mechanisms andFunctions", Prentice-Hall, Englewood Cliffs, N.J. (1976); Essays Med.Biochem., 2, 1-35 (1976); Hospital Practice, 12, 33-43 (1977);Perturbation of Complement in Disease, Chap. 15 in BiologicalAmplification Systems in Immunology (Ed. Day and Good), Plenum, New Yorkand London (1977); Am. J. Clin. Pathology, 68, 647-659 (1977).

The complement system can be considered to consist of three sub-systems:(1) a recognition unit (C1q) which enables it to combine with antibodymolecules that have detected a foreign invader; (2) an activation unit(C1r, C1s, C2, C4, C3) which prepares a site on the neighboringmembrane; and (3) an attack unit (C5, C6, C7, C8 and C9) which creates a"hole" in the membrane. The membrane attack unit is non-specific; itdestroys invaders only because it is generated in their neighborhood. Inorder to minimize damage to the host's own cells, its activity must belimited in time. This limitation is accomplished partly by thespontaneous decay of activated complement and partly by interference byinhibitors and destructive enzymes. The control of complement, however,is not perfect, and there are times when damage is done to the host'scells. Immunity is, therefore, a double-edged sword.

Activation of the complement system also accelerates blood clotting.This action comes about by way of the complement-mediated release of aclotting factor from platelets. The biologically active complementfragments and complexes can become involved in reactions that damage thehost's cells, and these pathogenic reactions can result in thedevelopment of immune-complex diseases. For example, in some forms ofnephritis, complement damages the basal membrane of the kidney,resulting in the escape of protein from the blood into the urine. Thedisease disseminated lupus erythematosus belongs in this category; itssymptoms include nephritis, visceral lesions and skin eruptions. Thetreatment of diphtheria or tetanus with the injection of large amountsof antitoxin sometimes results in serum sickness, an immune-complexdisease. Rheumatoid arthritis also involves immune complexes. Likedisseminated lupus erythematosus, it is an autoimmune disease in whichthe disease symptoms are caused by pathological effects of the immunesystem in the host's tissues. In summary, the complement system has beenshown to be involved with inflammation, coagulation, fibrinolysis,antibody-antigen reactions and other metabolic processes.

In the presence of antibody-antigen complexes the complement proteinsare involved in a series of reactions which may lead to irreversiblemembrane damage if they occur in the vicinity of biological membranes.Thus, while complement constitutes a part of the body's defensemechanism against infection it also results in inflammation and tissuedamage in the immunopathological process. The nature of certain of thecomplement proteins, suggestions regarding the mode of complementbinding to biological membranes and the manner in which complementeffects membrane damage are discussed in Annual Review in Biochemistry,38, 389 (1969); Journal of Immunology, 119, 1-8, 1195, 1358-1364, 1482(1977).

A variety of substances have been disclosed as inhibiting the complementsystem, i.e., as complement inhibitors. For example, the compounds3,3'-ureylenebis[6-(2-amino-8-hydroxy-6-sulfo-1-naphthylazo)benzenesulfonicacid], tetrasodium salt (chlorazol fast pink), heparin and a sulphateddextran have been reported to have an anticomplementary effect, BritishJournal of Experimental Pathology, 33, 327-339 (1952). German Pat. No.2,254,893 or South African Pat. No. 727,923 discloses certain1-(diphenylmethyl)-4-(3-phenylallyl)piperazines useful as complementinhibitors. Other chemical compounds having complement inhibitingactivity are disclosed in, for example, Journal of Medicinal Chemistry,12, 415-419; 902-905; 1049-1052; 1053-1056 (1969); Canadian Journal ofBiochemistry, 47, 547-552 (1969); The Journal of Immunology, 104,279-288 (1970); The Journal of Immunology, 106, 241-245 (1971); TheJournal of Immunology, 111, 1061-1066 (1973); Biochim. Biophys. Acta,317, 539-548 (1973); Life Sciences, 13, 351-362 (1973); Journal ofImmunology, 113, 584 (1974); Immunology, 26, 819-829 (1974); Journal ofMedicinal Chemistry, 17, 1160-1167 (1974); Biochim. Biophys. Res. Comm.,67, 225-263 (1975); Ann. N.Y. Acad. Sci., 256, 441-450 (1975); Journalof Medicinal Chemistry, 19, 634-639, 1079 (1976); Journal of Immunology,118, 466 (1977); Arch. Int. Pharmacodyn., 226, 281-285 (1977); Biochem.Pharmacol. 26, 325-329 (1977); Journal Pharm. Sci., 66, 1367-1377(1977); Chem. Pharm. Bull., 25, 1202-1208 (1977); Biochim. Biophys.Acta, 484, 417-422 (1977) and Journal Clin. Microbiology, 5, 278-284(1977).

It has been reported that the known complement inhibitorsepsilon-aminocaproic acid and tranexamic acid have been used withsuccess in the treatment of hereditary angioneurotic edema, a diseasestate resulting from an inherited deficiency or lack of function of theserum inhibitor of the activated first component of complement (C1inhibitor), The New England Journal of Medicine, 286, 808-812 (1972),287, 452-454 (1972); Ann. Intern. Med., 84, 580-593 (1976); J. Allergyand Clin. Immunology, 60, 38-40 (1977).

It has also been reported that the drug pentosan-polysulfoester has ananticomplementary activity on human serum, both in vitro and in vivo, asjudged by the reduction in total hemolytic complement activity;Pathologie Biologie, 25, 33-36, 25 (2), 105-108, 25 (3), 179-184 (1977).

Publications related to the biological use of Suramin compounds for thepurpose of inhibiting the complement system, including humans, asdetermined by the in vivo and in vitro testing of the blood serum ofwarm-blooded animals are:

B. Stuber and K. Lang, Arch. Exptl. Path. Pharmacol., 154, 41-49 (1930)[C. A. 25, 3067 (1931)];

F. Klopstock, Zeitschrift fur Immunitatsforschung und experimentalleTherapie, 75, 348-354 (1932);

H. J. Schmid, Schweiz. Med. Woch., 96, 1267-1269 (1966);

K. Lauenstein, Bayer-Symposium I, 25-30 (1969);

J. S. C. Fong and R. A. Good, Clin. Exp. Immunol., 10, 127-138 (1972);

V. Eisen and C. Loveday, Br. J. Pharmac., 49, 678-687 (1973);

D. Brackertz and F. Kueppers, Allergol. Et Immunopath., 11, 163-168(1974);

E. Raepple, H-U. Hill and M. Loos, Immunochemistry, 13 (3), 251-255(1976).

SUMMARY OF THE INVENTION

This invention is concerned with certainnaphthalenetetrayltetrakis(sulfonylimino)-aryl multicarboxylic acids andsalts thereof, and their utility of inhibiting complement activity inbody fluids.

This invention is particularly concerned with compounds havingcomplement inhibiting activity of the general formula (I): ##STR1##wherein R₁ is COOR₃, wherein R₃ is selected from the group consisting ofhydrogen and a pharmaceutically acceptable salt cation; and R₂ isselected from the group consisting of hydrogen and COOR₃.

Of particular interest is the group of compounds encompassed by theabove general structure (I), and illustrated by the structures (II) and(III): ##STR2## wherein R₄ is selected from the group consisting ofhydrogen and a pharmaceutically acceptable salt cation. Operablepharmaceutically acceptable salts include, for example, those of alkalimetal, alkaline earth metal, ammonia and substituted ammonia, such astrialkylamines (C₁ -C₆), piperidine, pyrazine, cycloalkylamines (C₄ -C₈)and alkanolamines (C₂ -C₆).

Representative compounds encompassed within this invention include, forexample,5,5',5",5'"-[1,3,5,7-naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,2,3-benzenetricarboxylicacid, dodecasodium salt;5,5',5",5'"-[1,3,6,7-naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,2,3-benzenetricarboxylicacid, dodecasodium salt;5,5',5",5'"-[1,3,6,8-naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,2,3-benzenetricarboxylicacid, dodecasodium salt;5,5',5",5'"-[1,3,5,7-naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,3-benzenedicarboxylicacid, octasodium salt;5,5',5",5'"-[1,3,6,7-naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,3-benzenedicarboxylicacid, octasodium salt; and,5,5',5"5'"-[1,3,6,8-naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,3-benzenedicarboxylicacid, octasodium salt.

The compounds of this invention may be prepared, for example, accordingto the following illustrative general procedure: Treatment of 1,3,5(or6),7(or 8)-naphthalenetetrasulfonic acid, tetrasodium salt with thionylchloride provides 1,3,5-(or 6),7(or 8)-naphthalenetetrasulfonylchloride. Heating the latter in pyridine with5-amino-1,2,3-benzenetricarboxylic acid, triphenyl ester provides5,5',5",5'"-[1,3,5(or 6),7(or8)-naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,2,3-benzenetricarboxylicacid, dodecaphenyl ester. Saponification with 2 N sodium hydroxide atroom temperature followed by neutralization with acetic acid yields5,5',5",5'"-[1,3,5(or 6),7(or8)-naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,2,3-benzenetricarboxylicacid, dodecasodium salt.

Condensation of 1,3,5(or 6),7(or 8)-naphthalenetetrasulfonyl chloridewith 5-amino-1,3-benzenedicarboxylic, bis(2-methoxyethyl)ester produces5,5',5",5'"-[1,3,5(or 6),7(or8)-naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,3-benzenedicarboxylicacid, octakis(2-methoxyethyl)ester. Saponification with sodium hydroxidegives 5,5',5",5'"-[1,3,5(or 6),7(or8)-naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,3-benzenedicarboxylicacid, octasodium salt.

This invention is also concerned with a method of inhibiting thecomplement system in a body fluid, such as blood serum, which comprisessubjecting the body fluid complement to the action of an effectivecomplement inhibiting amount of a compound of this invention. The methodof use aspect of this invention is further concerned with a method ofinhibiting the complement system in a warm-blooded animal whichcomprises administering to said animal an effective complementinhibiting amount of a compound of this invention.

The compounds of this invention find utility as complement inhibitors inbody fluids such as blood, plasma, serum, synovial fluid, cerebrospinalfluid, or pathological accumulations of fluid such as pleural effusion.As such, they may be used to ameliorate or prevent those pathologicalreactions requiring the function of complement and in the therapeutictreatment of warm-blooded animals having immunologic diseases such asrheumatoid arthritis, systemic lupus erythematosus, certain kinds ofglomerulonephritis, certain kinds of auto-allergic hemolytic anemia,certain kinds of platelet disorders and certain kinds of vasculitis.

These compounds may also be used in the treatment of warm-bloodedanimals having non-immunologic diseases such as paroxysmal nocturnalhemoglobinuria, hereditary angioneurotic edema and inflammatory statesinduced by the action of bacterial or lysosomal enzymes on theappropriate complement components as for example, inflammation followingcoronary occlusion. They may also be useful in the treatment oftransplant rejection and as blood culture and transport mediums.

DETAILED DESCRIPTION OF THE INVENTION

The following examples describe in detail the preparation andformulation of representative compounds of the present invention.

EXAMPLE 1 1,3,5,7-Naphthalenetetrasulfonyl chloride

To 240 ml. of 30-33% fuming sulfuric acid at 120° C. is added,portionwise, 60 g. of 1,5-naphthalenedisulfonic acid, disodium salt. Themixture is heated at 180°-190° C. for 23 hours. The solution is cooled,allowed to crystallize and filtered through a coarse, sintered glassfunnel. The product is washed with concentrated sulfuric acid to give1,3,5,7-naphthalenetetrasulfonic acid as an off-white pasty solid.Without further purification, the solid is added, portionwise, to 150ml. of chlorosulfonic acid and the mixture is heated at 145°-150° C. for1-5 hours. The mixture is cooled to room temperature and filtered. Theproduct is washed, in order, with chlorosulfonic acid, methylenechloride, acetonitrile, water, acetonitrile and finally with ether. Theproduct is air-dried and then dried further in an Abderhalden apparatusat 76° C. to provide 33.9 g. of the desired product as a colorlesspowder, mp. 263°-270° C.

EXAMPLE 2 5,5',5",5'"-[1,3,5,7-Naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,3-benzenedicarboxylicacid, octakis(2-methoxyethyl)ester

To a solution of 23.78 g. of 5-amino-1,3-benzenedicarboxylic acid,bis(2-methoxyethyl)ester in 100 ml. of acetonitrile and 6.52 ml. ofpyridine (both dried over molecular sieves) is added 10.4 g. of1,3,5,7-naphthalenetetrasulfonyl chloride. The mixture is stirred atroom temperature for 1.5 hours and then at 50°-60° C. for one hour. Thesolution is poured into water and the mixture is filtered throughdiatomaceous earth. The oily product is dissolved in methylene chlorideand the solution is washed with 2 N hydrochloric acid, then with brineand the organic phase is dried over anhydrous sodium sulfate. The driedsolution is evaporated to a pink powder which is chromatographed on asilica gel column using 30-50% acetone plus 1% acetic acid in hexane asthe eluant. The product fractions are evaporated and the residue isdissolved in 100 ml. of boiling tetrahydrofuran and filtered throughdiatomaceous earth. The filtrate is diluted with 400 ml. of ether toprecipitate the product which is filtered and dried overnight in anAbderhalden apparatus at 110° C. to provide the desired product, 13.05g. of colorless powder, mp. 210°-212° C.

EXAMPLE35,5',5",5'"-[1,3,5,7-Naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,3-benzenedicarboxylicacid, octasodium salt

A mixture of 12.52 g. of5,5',5",5'"-[1,3,5,7-naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,3-benzenedicarboxylicacid, octakis(2-methoxyethyl)ester and 60 ml. of 2 N sodium hydroxide isstirred at room temperature for one hour. Acetic acid (3.67 g.) is addedand the solution is diluted with 375 ml. of absolute ethanol. Themixture is filtered and the product is washed with ethanol and then withether. After air drying, the product is dried overnight in anAbderhalden apparatus at 110° C. to provide 10.93 g. of the desiredproduct as an orange powder.

EXAMPLE 45,5',5",5'"-[1,3,5,7-Naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,2,3-benzenetricarboxylicacid, dodecaphenyl ester

To a solution of 36.27 g. of 5-amino-1,2,3-benzenetricarboxylic acid,triphenyl ester in 150 ml. of dry pyridine is added 12.0 g. of1,3,5,7-naphthalenetetrasulfonyl chloride and the mixture is stirred atroom temperature for one hour and then at 55°-75° C. for 2 hours. Thesolution is poured into 840 ml. of 2 N hydrochloric acid and theresulting mixture is extracted with methylene chloride. The aqueousphase is filtered to remove the suspended solid which is washed withwater and methylene chloride to provide 32.4 g. of a cream powder. Theproduct is crystallized from acetone-methylene chloride and is thenrefluxed in acetonitrile:methanol (2:1) solution. The product isfiltered and the filtrate cooled to 5° C. to provide additional product.The combined fractions are dried overnight in an Abderhalden apparatusat 110° C. to give 21.45 g. of beige powder, mp. >300° C.

EXAMPLE 55,5',5",5'"-[1,3,5,7-Naphthalenetetrayltetrakis[sulfonylimino)]-tetra-1,2,3-benzenetricarboxylicacid, dodecasodium salt

A mixture of 10.95 g. of5,5',5",5'"-[1,3,5,7-naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,2,3-benzenetricarboxylicacid, dodecaphenyl ester and 85 ml. of 2 N sodium hydroxide is stirredat room temperature for one hour. The solution is neutralized with 6.29ml. of acetic acid and concentrated in vacuo to approximately 65 ml. Thesolution is poured into 550 ml. of absolute ethanol, filtered and theproduct washed with ethanol and ether. The product is dissolved in 65ml. of water, filtered, and adjusted to pH 7.4 with 1 N sodiumhydroxide. The solution is poured into 750 ml. of absolute ethanol,sodium acetate trihydrate being added to improve coagulation of theprecipitate. The gummy precipitate is triturated over fresh ethanoluntil it solidifies and the mixture is filtered. The solid is taken upin water again and re-adjusted to pH 7.4 with acetic acid. The solutionis poured into 1.5 liters of absolute ethanol, 5 g. of sodium acetatetrihydrate is added and the mixture is stirred for 2 hours. The mixtureis filtered and the product is washed with ethanol and ether, and isdried overnight at 110° C. in an Abderhalden apparatus to give 8.2 g. ofthe desired product as an orange powder.

EXAMPLE 6 Preparation of Compressed Tablet

    ______________________________________                                        Ingredient              mg/Tablet                                             ______________________________________                                        Active Compound         0.5-500                                               Dibasic Calcium Phosphate N.F.                                                                        qs                                                    Starch USP              40                                                    Modified Starch         10                                                    Magnesium Stearate USP  1-5                                                   ______________________________________                                    

EXAMPLE 7 Preparation of Compressed Tablet--Sustained Action

    ______________________________________                                        Ingredient          mg/Tablet                                                 ______________________________________                                        Active Compound as Aluminum                                                                       0.5-500 (as acid                                          Lake*, Micronized   equivalent)                                               Dibasic Calcium Phosphate N.F.                                                                    qs                                                        Alginic Acid        20                                                        Starch USP          35                                                        Magnesium Stearate USP                                                                            1-10                                                      ______________________________________                                         *Complement inhibitor plus aluminum sulfate yields aluminum complement        inhibitor. Complement inhibitor content in aluminum lake ranges from          5-30%.                                                                   

EXAMPLE 8 Preparation of Hard Shell Capsule

    ______________________________________                                        Ingredient              mg/Capsule                                            ______________________________________                                        Active Compound       0.5-500                                                 Lactose, Spray Dried  qs                                                      Magnesium Stearate    1-10                                                    ______________________________________                                    

EXAMPLE 9 Preparation of Oral Liquid (Syrup)

    ______________________________________                                        Ingredient        % W/V                                                       ______________________________________                                        Active Compound   0.05-5                                                      Liquid Sugar      75.0                                                        Methyl Paraben USP                                                                              0.18                                                        Propyl Paraben USP                                                                              0.02                                                        Flavoring Agent   qs                                                          Purified Water qs ad.                                                                           100.0                                                       ______________________________________                                    

EXAMPLE 10 Preparation of Oral Liquid (Elixir)

    ______________________________________                                        Ingredient        % W/V                                                       ______________________________________                                        Active Compound   0.05-5                                                      Alcohol USP       12.5                                                        Glycerin USP      45.0                                                        Syrup USP         20.0                                                        Flavoring Agent   qs                                                          Purified Water qs ad                                                                            100.0                                                       ______________________________________                                    

EXAMPLE 11 Preparation of Oral Suspension (Syrup)

    ______________________________________                                        Ingredient            % W/V                                                   ______________________________________                                        Active Compound as Aluminum                                                                         0.05-5                                                  Lake, Micronized      (acid equivalent)                                       Polysorbate 80 USP    0.1                                                     Magnesium Aluminum Silicate,                                                  Colloidal             0.3                                                     Flavoring Agent       qs                                                      Methyl Paraben USP    0.18                                                    Propyl Paraben USP    0.02                                                    Liquid Sugar          75.0                                                    Purified Water qs ad  100.0                                                   ______________________________________                                    

EXAMPLE 12 Preparation of Injectable Solution

    ______________________________________                                        Ingredient        % W/V                                                       ______________________________________                                        Active Compound   0.05-5                                                      Benzyl Alcohol N.F.                                                                             0.9                                                         Water for Injection                                                                             100.0                                                       ______________________________________                                    

EXAMPLE 13 Preparation of Injectable Oil

    ______________________________________                                        Ingredient      % W/V                                                         ______________________________________                                        Active Compound 0.05-5                                                        Benzyl Alcohol  1.5                                                           Sesame Oil qs ad                                                                              100.0                                                         ______________________________________                                    

EXAMPLE 14 Preparation of Intra-Articular Product

    ______________________________________                                        Ingredient               Amount                                               ______________________________________                                        Active Compound          2-20 mg                                              NaCl (physiological saline)                                                                            0.9%                                                 Benzyl Alcohol           0.9%                                                 Sodium Carboxymethylcellulose                                                                          1-5%                                                 pH adjusted to 5.0--Water for Injection qs ad                                                          100%                                                 ______________________________________                                    

EXAMPLE 15 Preparation of Injectable Depo Suspension

    ______________________________________                                        Ingredient            % W/V                                                   ______________________________________                                        Active Compound       0.05-5                                                                        (acid equivalent)                                       Polysorbate 80 USP    0.2                                                     Polyethylene Glycol 4000 USP                                                                        3.0                                                     Sodium Chloride USP   0.8                                                     Benzyl Alcohol N.F.   0.9                                                     HCl to pH 6-8         qs                                                      Water for Injection qs ad                                                                           100.0                                                   ______________________________________                                    

EXAMPLE 16 Preparation of Dental Paste

    ______________________________________                                        Ingredient           % W/W                                                    ______________________________________                                        Active Compound      0.05-5                                                   Zinc Oxide           15                                                       Polyethylene Glycol 4000 USP                                                                       50                                                       Distilled Water qs   100                                                      ______________________________________                                    

EXAMPLE 17 Preparation of Dental Ointment

    ______________________________________                                        Ingredient         % W/W                                                      ______________________________________                                        Active Compound    0.05-5                                                     Petrolatum, White USP qs                                                                         100                                                        ______________________________________                                    

EXAMPLE 18 Preparation of Dental Cream

    ______________________________________                                        Ingredient          % W/W                                                     ______________________________________                                        Active Compound     0.05-5                                                    Mineral Oil         50                                                        Beeswax             15                                                        Sorbitan Monostearate                                                                             2                                                         Polyoxyethylene 20 Sorbitan                                                   Monostearate        3                                                         Methyl Paraben USP  0.18                                                      Propyl Paraben USP  0.02                                                      Distilled Water qs  100                                                       ______________________________________                                    

EXAMPLE 19 Preparation of Topical Cream

    ______________________________________                                        Ingredient        % W/W                                                       ______________________________________                                        Active Compound   0.05-5                                                      Sodium Lauryl Sulfate                                                                           1                                                           Propylene Glycol  12                                                          Stearyl Alcohol   25                                                          Petrolatum, White USP                                                                           25                                                          Methyl Paraben USP                                                                              0.18                                                        Propyl Paraben USP                                                                              0.02                                                        Purified Water qs 100                                                         ______________________________________                                    

EXAMPLE 20 Preparation of Topical Ointment

    ______________________________________                                        Ingredient         % W/W                                                      ______________________________________                                        Active Compound    0.05-5                                                     Cholesterol        3                                                          Stearyl Alcohol    3                                                          White Wax          8                                                          Petrolatum, White USP qs                                                                         100                                                        ______________________________________                                    

EXAMPLE 21 Preparation of Spray Lotion (non-Aerosol)

    ______________________________________                                        Ingredient         % W/W                                                      ______________________________________                                        Active Compound    0.05-5                                                     Isopropyl Myristate                                                                              20                                                         Alcohol (Denatured) qs                                                                           100                                                        ______________________________________                                    

EXAMPLE 22 Preparation of Buccal Tablet

    ______________________________________                                        Ingredient          g/Tablet                                                  ______________________________________                                        Active Ingredient   0.00325                                                   6 × Sugar     0.29060                                                   Acacia              0.01453                                                   Soluble Starch      0.01453                                                   F. D. & C. Yellow No. 6 Dye                                                                       0.00049                                                   Magnesium Stearate  0.00160                                                                       0.32500                                                   ______________________________________                                    

The final tablet will weigh about 325 mg. and may be compressed intobuccal tablets in flat faced or any other tooling shape convenient forbuccal administration.

EXAMPLE 23 Preparation of Lozenge

    ______________________________________                                        Ingredient           g/Lozenge                                                ______________________________________                                        Active Ingredient    0.0140                                                   Kompact® Sugar (Sucrest Co.)                                                                   0.7138                                                   6 × Sugar      0.4802                                                   Sorbitol (USP Crystalline)                                                                         0.1038                                                   Flavor               0.0840                                                   Magnesium Stearate   0.0021                                                   Dye                  qs                                                       Stearic Acid         0.0021                                                                        1.4000                                                   ______________________________________                                    

The ingredients are compressed into 5/8" flat based lozenge tooling.Other shapes may also be utilized.

The compounds of the present invention may be administered internally,e.g., orally or parenterally, e.g., intra-articularly, to a warm-bloodedanimal to inhibit complement in the body fluid of the animal, suchinhibition being useful in the amelioration or prevention of thosereactions dependent upon the function of complement, such asinflammatory process and cell membrane damage induced byantigen-antibody complexes. A range of doses may be employed dependingon the mode of administration, the condition being treated and theparticular compound being used. For example, for intravenous orsubcutaneous use from about 5 to about 50 mg./kg./day, or every sixhours for more rapidly excreted salts, may be used. For intra-articularuse for large joints such as the knee, from about 2 to about 20mg./joint per week may be used, with proportionally smaller doses forsmaller joints. The dosage range is to be adjusted to provide optimumtherapeutic response in the warm-blooded animal being treated. Ingeneral, the amount of compound administered can vary over a wide rangeto provide from about 5 mg./kg. to about 100 mg./kg. of body weight ofanimal per day. The usual daily dosage for a 70 kg. subject may varyfrom about 350 mg. to about 3.5 g. Unit doses of the acid or salt cancontain from about 0.5 mg. to about 500 mg.

While in general the sodium salts of the acids of the invention aresuitable for parenteral use, other salts may also be prepared, such asthose of primary amines, e.g., ethylamine; secondary amines, e.g.,diethylamine or diethanol-amine; tertiary amines, e.g., pyridine ortriethylamine or 2-dimethylaminomethyldibenzofuran; aliphatic diamines,e.g., decamethylenediamine; and aromatic diamines, can be prepared. Someof these are soluble in water, others are soluble in saline solution,and still others are insoluble and can be used for purposes of preparingsuspensions for injection. Furthermore, as well as the sodium salt,those of the alkali metals, such as potassium and lithium; of ammonia;and of the alkaline earth metals, such as calcium or magnesium, may beemployed. It will be apparent, therefore, that these salts embrace, ingeneral, derivatives of salt-forming cations.

The compounds of the present invention may also be administeredtopically in the form of ointments, creams, lotions and the like,suitable for the treatment of complement dependent dermatologicaldisorders.

Moreover, the compounds of the present invention may be administered inthe form of dental pastes, ointments, buccal tablets and othercompositions suitable for application periodontally for the treatment ofperiodontitis and related diseases of the oral cavity.

In therapeutic use, the compounds of this invention may be administeredin the form of conventional pharmaceutical compositions. Suchcompositions may be formulated so as to be suitable for oral orparenteral administration. The active ingredient may be combined inadmixture with a pharmaceutically acceptable carrier, which carrier maytake a wide variety of forms depending on the form of preparationdesired for administration, i.e., oral or parenteral. The compounds canbe used in compositions such as tablets. Here, the principal activeingredient is mixed with conventional tabletting ingredients such ascorn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesiumstearate, dicalcium phosphate, gums, or similar materials as non-toxicpharmaceutically acceptable diluents or carriers. The tablets or pillsof the novel compositions can be laminated or otherwise compounded toprovide a dosage form affording the advantage of prolonged or delayedaction or predetermined successive action of the enclosed medication.For example, the tablet or pill can comprise an inner dosage and anouter dosage component, the latter being in the form of an envelope overthe former. The two components can be separated by an enteric layerwhich serves to resist disintegration in the stomach and permits theinner component to pass intact into the duodenum or to be delayed inrelease. A variety of materials can be used for such enteric layers orcoatings, such materials including a number of polymeric acids ormixtures of polymeric acids with such materials as shellac, shellac andcetyl alcohol, cellulose acetate and the like. A particularlyadvantageous enteric coating comprises a styrene maleic acid copolymertogether with known materials contributing to the enteric properties ofthe coating. The tablet or pill may be colored through the use of anappropriate non-toxic dye, so as to provide a pleasing appearance.

The liquid forms in which the novel compositions of the presentinvention may be incorporated for administration include suitableflavored emulsions with edible oils, such as, cottonseed oil, sesameoil, coconut oil, peanut oil, and the like, as well as elixirs andsimilar pharmaceutical vehicles. Sterile suspensions or solutions can beprepared for parenteral use. Isotonic preparations containing suitablepreservatives are also desirable for injection use.

The term dosage form, as described herein, refers to physically discreteunits suitable as unitary dosage for warm-blooded animal subjects, eachunit containing a predetermined quantity of active component calculatedto produce the desired therapeutic effect in association with therequired pharmaceutical diluent, carrier or vehicle. The specificationfor the novel dosage forms of this invention are indicated bycharacteristics of the active component and the particular therapeuticeffect to be achieved or the limitations inherent in the art ofcompounding such an active component for therapeutic use in warm-bloodedanimals as disclosed in this specification. Examples of suitable oraldosage forms in accord with this invention are tablets, capsules, pills,powder packets, granules, wafers, cachets, teaspoonfuls, dropperfuls,ampules, vials, segregated multiples of any of the foregoing and otherforms as herein described.

The complement inhibiting activity of the compounds of this inventionhas been demonstrated by one or more of the following identified tests:(i) Test Code 026 (C1 inhibitor): This test measures the ability ofactivated human C1 to destroy fluid phase human C2 in the presence of C4and appropriate dilutions of the test compound. An active inhibitorprotects C2 from C1 and C4; (ii) Test Code 035 (C3-C9 inhibitor): Thistest determines the ability of the late components of human complement(C3-C9) to lyse EAC 142 in the presence of appropriate dilutions of thetest compound. An active inhibitor protects EAC 142 from lysis by humanC3-C9; (iii) Test Code 036 (C-Shunt inhibitor): In this test humanerythrocytes rendered fragile are lysed in autologous serum via theshunt pathway activated by cobra venom factor in the presence ofappropriate dilutions of the test compound. Inhibition of the shuntpathway results in failure of lysis; (iv) Forssman Vasculitis Test:Here, the well known complement dependent lesion, Forssman vasculitis,is produced in guinea pigs by intradermal injection of rabbitanti-Forssman antiserum. The lesion is measured in terms of diameter,edema and hemorrhage and the extent to which a combined index of theseis inhibited by prior intraperitoneal injection of the test compound at200 mg./kg. is then reported, unless otherwise stated; (v) ForssmanShock Test: Lethal shock is produced in guinea pigs by an i.v. injectionof anti-Forssman antiserum and the harmonic mean death time of treatedguinea pigs is compared with that of simultaneous controls; (vi)Complement Level Reduction Test: In this test, the above dosed guineapigs, or others, are bled for serum and the complement level isdetermined in undiluted serum by the capillary tube method of U.S. Pat.No. 3,876,376 and compared to undosed control guinea pigs; (vii) Cap 50Test: Here, appropriate amounts of the test compound are added to a poolof guinea pig serum in vitro, after which the undiluted serum capillarytube assay referred to above is run. The concentration of compoundinhibiting 50% is reported; (viii) Guinea Pig Intraperitoneal Test(GPIP): Guinea pigs weighing about 300 g. are dosed intraperitoneally(i.p.) with 200 mg./kg. of the test compound dissolved in saline andadjusted to pH 7-8. Approximately 0.4 ml. blood samples, taken byorbital sinus puncture 30 minutes and one hour after injections, arecollected directly into centrifuge tubes; 5 ml. blood samples, taken bydecapitation 2 hours after injection are collected directly into diSPo®beakers. The samples were allowed to clot, centrifuged, and theresultant sera are assayed for complement activity using the capillarycomplement assay. Percent inhibition is calculated by comparison withsimultaneous controls. The results appear in Table I together withresults of Test Code 026, 035, 036, Cap 50 and % Inhibition. Table Ishows that the principal compound of the invention possesses highlysignificant in vitro and in vivo complement inhibiting activity inwarm-blooded animals.

                                      TABLE I                                     __________________________________________________________________________    BIOLOGICAL ACTIVITIES                                                                                                 In Vivo Activity                                                              (Guinea Pig)                                                                  % Inhibition                                             Cl  C-Late                                                                            Shunt Inhibition                                                                           Intraperitoneal                                          026*                                                                              035*                                                                              036*         Time (Minutes                         Compound           Wells                                                                             Wells                                                                             Wells   Cap 50*                                                                            30 60 120                             __________________________________________________________________________    5,5', 5", 5'"-[1,3,5,7-Naphthalenetetra-                                      yltetrakis(sulfonylimino)]-tetra-1,3-                                                            +7**                                                                              +2  +6      58   -66                                                                              -71                                                                              -76                             benzenediacrboxylic acid, octasodium                                          salt                                                                          5,5', 5", 5'"-[1,3,5,7-Naphthalenetetra-                                      yltetrakis(sulfonylimino)1-tetra-                                                                +2  +4  +6      96   -80                                                                              -77                                                                              -85                             1,2,3-benzenetricarboxylic acid,                                              dodecasodium salt                                                             __________________________________________________________________________     *Code designation for tests employed as referred herein.                      **Activity in wells, a serial dilution assay. Higher well number indicate     higher activity. The serial dilutions are twofold.                       

We claim:
 1. A compound of the formula: ##STR3## wherein R₁ is COOR₃ ;wherein R₃ is selected from the group consisting of hydrogen and apharmaceutically acceptable salt cation; and R₂ is selected from thegroup consisting of hydrogen and COOR₃.
 2. The compound according toclaim 1,5,5',5",5'"-[1,3,5,7-naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,2,3-benzenetricarboxylicacid, dodecasodium salt.
 3. The compound according to claim 1,5,5',5",5'"-[1,3,6,7-naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,2,3-benzenetricarboxylicacid, dodecasodium salt.
 4. The compound according to claim 1,5,5',5",5'"-[1,3,6,8-naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,2,3-benzenetricarboxylicacid, dodecasodium salt.
 5. The compound according to claim 1,5,5',5",5'"-[1,3,5,7-naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,3-benzenedicarboxylicacid, octasodium salt.
 6. The compound according to claim 1,5,5',5",5'"-[1,3,6,7-naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,3-benzenedicarboxylicacid, octasodium salt.
 7. The compound according to claim 1,5,5',5",5'"-[1,3,6,8-naphthalenetetrayltetrakis(sulfonylimino)]-tetra-1,3-benzenedicarboxylicacid, octasodium salt.